Antibody-containing blend obtained from eggs

ABSTRACT

The invention relates to blending eggs from lots produced over the course of the egg laying cycle of immunized avians, to form an antibody-containing blend containing both early-stage antibodies and late-stage antibodies. The administration of an effective amount of such an antibody-containing blend to a subject increases the efficacy of the antibody blend its ability to bind antigen over that of a substance containing the same amount of antibody obtained from eggs containing either early-stage antibodies or late-stage antibodies only.

RELATED APPLICATIONS

[0001] This application is a continuation of U.S. patent application Ser. No. 09/056,690, filed Apr. 8, 1998 now issued as U.S. Pat. No. 6,217,865, on Apr. 14, 2001 and U.S. patent application Ser. No. 09/687,614 filed Oct. 16, 2000, now abandoned.

FIELD OF THE INVENTION

[0002] The present invention relates to a method of increasing the efficacy of an antibody's ability to bind to an antigen in a subject. More particularly, the invention relates to blending eggs from egg lots produced over the course of an avians egg laying cycle (i.e. 30-60 weeks of laying eggs) to form an antibody-containing substance. The administration of an effective amount of such an antibody-containing blend to a subject increases the efficacy of the antibody's ability to bind antigen due to the synergistic effect of early-stage and late-stage antibodies.

BACKGROUND OF THE INVENTION

[0003] The induction of an immune response in a living animal causes many things to occur within that animal, one of which is the production of antibodies. There are several characteristics of an antibody response including specificity, amount, isotype (class) and affinity of the antibodies produced. The specificity determines the ability of the antibody to distinguish one immunogen from another. The amount of antibody is a function of the number of responding B cells, their rate of antibody synthesis, and the persistence of the antibody after production. The persistence of the antibody in different tissues of the body is determined by its isotype, wherein each isotype has a different half-life in vivo. The isotypic composition of an antibody response also determines the biological functions these antibodies can perform and the sites in which they will be found. Finally, the strength of binding or affinity of the antibody to its antigen is important because the higher the affinity, the less the amount of antibody that will be necessary to eliminate the antigen.

[0004] These different characteristics of the antibodies occur at different times during the development of a mature antibody. This process is known as affinity maturation. The affinity maturation process varies not only in response to each individual immunogen, but also for every individual animal which is undergoing the immune response.

[0005] In recent years, it has been determined that avians and other animal species produce different immunoglobulins (Igs) than do mammals. In particular, mammals produce a unique series of Igs, including IgM, IgG, IgE and IgA among others. In the vertebrate classes of Amphibia, Reptilia and Aves, the primary Ig which is produced in the egg yolk is IgY. IgY is believed to be the evolutionary ancestor of IgG and IgE. (Warr et al., Immunology Today, Vol. 16, No. 8, pp 392-298, 1995).

[0006] It has been scientifically documented that antibodies produced by an animal's T-cells mature over time after exposure to an antigen. The maturation process is driven by mutational genetic drift and repertoire shift causing changes in antibody binding site structure (Roi H, et al., Immunology, 5^(th) Edition, pp. 112-153, 1998). These changes result in greater average affinity and faster binding kinetics of IgY antibody through clonal selection and are optimized for T-Cell activation within the animal's body. This may be different from optimization for inhibition of binding to other types of antigen receptors.

[0007] Passive Immunization

[0008] In the art, there is a process known as passive immunization, which relates to the transfer of immunity from one species to another. For example, in a common form of passive immunization, an avian is immunized with one or more immunogens. Immunization with such immunogens induces an immune response in the avian. Antibodies to the immunogens are produced by the avian during the immune response. A large number of these antibodies become concentrated in the egg of the avian. Administration to an animal of the egg containing such antibodies, or administration of the purified antibodies themselves, causes that animal to become passively immunized to the corresponding immunogens due to the transfer of the antibodies. Such a passive immunization process is the subject of U.S. Pat. No. 4,748,018, assigned to DCV, Inc. and incorporated herein by reference.

[0009] The process of passive immunization using antibodies produced in the egg of an egg-producing animal requires, of course, the collection of antibody-containing eggs from an immunized avian. In this process, it is known in the art that the highest titers of antibodies are found in eggs produced at the early-stages of the egg-laying cycle of an immunized avian. As time goes on after an immune response induction, antibody titers rise quickly, eventually peak and then fall off. The production of antibody titer in the eggs of an immunized avian has been charted to generally form a bell-shaped curve over the entire egg laying life cycle of the avian (see R. Schade et al., ATLA, 19, pp. 403-419, 1991). Antibody titers can be somewhat maintained by revaccination. Either way, it is generally the practice in the art to collect eggs soon after vaccination or revaccination (i.e. within the first few (1-5) weeks) in order to obtain eggs having the highest antibody titers.

[0010] The inventors of the present invention have determined that although early-stage antibodies provide the highest titer of antibody, collection of both early-stage and late-stage antibodies produces an antibody-containing blend that is more effective than just early-stage or late-stage antibody alone.

SUMMARY OF THE INVENTION

[0011] It is an object of this invention to increase the efficacy of an egg antibody's ability to bind antigen. The efficacy of the antibody is increased by taking all lots of eggs produced by an avian over at least the first 30 weeks following immunization, wherein all lots of eggs contain antibodies produced in response to the immunization, and blending together these lots. The blending of eggs collected over such an extended period of time results in a more efficacious antibody-containing product when administered to a subject animal in terms of binding antigen.

[0012] In particular, the invention is directed to a method for increasing binding efficacy of an antibody in a subject comprising:

[0013] A. inducing an immune response in an egg-producing animal with at least one immunogen;

[0014] B. collecting eggs from the egg-producing animal over a portion of the course of an egg laying cycle following induction of said immune response;

[0015] C. blending all of the collected eggs together to form an antibody-containing blend; and

[0016] D. administering to the subject a portion of the antibody-containing blend which contains an effective titer of said antibody.

[0017] The invention is finally directed to a composition comprising an antibody-containing blend obtained from eggs collected from a period comprising at least 30 weeks following immunization of an avian with an immunogen, wherein the antibody-containing blend comprises early stage antibodies and late stage antibodies.

DETAILED DESCRIPTION OF THE INVENTION

[0018] Definitions:

[0019] The term “egg or fraction thereof” means any whole egg (table, hyperimmunized or otherwise) or any product derived therefrom.

[0020] The term “egg product” refers to any substance which is produced from one or more eggs or fractions thereof.

[0021] The term “immune-egg or fraction thereof” means one or more whole eggs or any product derived therefrom, obtained from egg producing animals which have been immunized and/or maintained in an immune state.

[0022] The term “antibody-containing blend” means any product that contains early-stage and late-stage antibodies produced by an immunized avian over a portion of the immunized avian's egg-laying cycle.

[0023] The term “egg-laying cycle” means the entire time span over which an egg-producing animal lays eggs prior to molting.

[0024] The term “egg lot” means a mixture of eggs which are collected from one or more egg-producing animals over a specific time-period after induction of an immune response, wherein the time-period is less than the entire egg-laying cycle.

[0025] The term “blended lot of eggs” means a mixture of more than one egg lots.

[0026] The term “blended egg product” means egg product derived from a blended lot of eggs.

[0027] The term “early-stage antibody” means any antibody produced by an animal during an immune response prior to the decline phase of antibody production after a secondary immune challenge, including clones produced during the log phase. Early-stage antibody being generally understood by those having skill in the art to possess lower affinity and slower binding kinetics than late-stage antibody.

[0028] The term “late-stage antibody” means any antibody produced by an animal during an immune response during only the decline phase of antibody production. Late-stage antibody being generally understood by those having skill in the art to posses higher affinity and faster binding kinetics than early-stage antibody

[0029] The term “effective amount of egg product” means an egg product containing an effective antibody titer. In particular, the amount of antibody is effective when the egg product is administered to a subject animal.

[0030] The term “hyperimmunization” means exposure to one or more antigens such that an immune response is elevated and maintained above the natural unexposed state.

[0031] The term “genetic vaccine” refers to a nucleic acid vaccine which is generally produced by recombinant technologies and which may elicit an immune response.

[0032] The term “administer” means any method of providing a subject with a substance, including orally, intranasally, parenterally (intravenously, intramuscularly, or subcutaneously), rectally or topically.

[0033] The term “animal” means any animal within the animal kingdom.

[0034] The term “target animal” refers to an animal which is immunized in order to produce an antibody-containing substance.

[0035] The term “subject animal” refers to the animal which is administered the antibody-containing substance.

[0036] The Invention

[0037] The present invention is an antibody-containing blend, and method for using the same, which comprises an effective amount of egg product obtained from a blended lot of eggs obtained from avians over a portion of the egg-laying cycle of the avians following immunization with at least one immunogen. The administration of an effective amount of such an antibody-containing blend to a subject animal increases the efficacy of the binding of the antibody to the antigen.

[0038] Due to the well known fact that later formed antibodies have greater affinity and faster binding kinetics, the inventors initially assumed that eggs produced later in the egg laying cycle and thus containing such late-stage antibodies, would bind antigen most effectively. However, to the inventors' surprise, antibody from eggs collected during the weeks 15-30 following immunization (i.e. late-stage antibodies) was only equivalently effective to the same concentration of antibody collected from eggs produced during weeks 1-15 (i.e. early-stage antibodies) more surprising has was the finding that neither early-stage antibodies alone or late-stage antibodies alone as effective as that seen from an equivalent amount of antibody from eggs collected over all of weeks 1-30 (i.e. early-stage antibodies in combination with late-stage antibodies).

[0039] As such, the surprising aspect of this invention is that antibody obtained from a blend of eggs covering the entire time span of at least the first 30 weeks of the egg-laying cycle following immunization, is much more efficacious in terms of binding antigen than antibody collected from one collection period of the egg-laying cycle.

[0040] Early-stage antibodies, having lower affinity with slower binding kinetics and derived earlier in the maturation process when combined with late-stage antibodies, having high affinity with faster binding kinetics, seem to produce a synergistic effect resulting in more efficient inhibition of binding of antigens to other types of receptors. Although early-stage antibodies possess non-optimal affinity and kinetics, these antibodies may bind to a greater variety of antigenic sites that better interfere with the antigen's binding to other receptors (i.e. more steric hindrance). Higher affinity late-stage IgY antibody also possess greater cross reactivity to proteins, peptides, or other structures that are similar to, but not part of the antigen of interest. This cross reactivity may effectively slow the binding kinetics of late-stage antibody to the antigen of interest, resulting in apparent weaker performance in inhibiting the antigen of interest's binding to other receptors. This effect substantiates reduced performance of late-stage antibody. Combination of early-stage and late-stage antibodies also results in early-stage antibodies partially inhibiting the cross reactive binding of late-stage antibody, allowing more opportunity for the higher affinity antibodies to bind to the antigen of interest and participate in blocking receptor binding. These synergistic effects produced by combining early-stage antibodies with steric hindrance advantages with late-stage antibodies possessing affinity/binding kinetic advantages result in overall performance gain in blended product vs. either early or late-stage product alone.

[0041] Immune-Egg Production

[0042] The immune egg can be produced by any egg-producing animal. It is preferred that the animal be a member of the class Aves or, in other words, an avian. Within the class Aves, fowl, and even more preferably, domesticated fowl, are preferred, but other members of this class, such as turkeys, ducks, and geese, are a suitable source of an immune egg.

[0043] When such egg-producing animals are brought to a specific state of immunization by means of, for example, periodic administrations of antigens, the animals will produce eggs which contain antibodies against those antigens.

[0044] Any immunogen or combination of immunogens may be employed as a vaccine. The immunogens can be antigens and can comprise proteins, enzymes, bacteria, viruses, protozoa, fungi, cellular antigens, allergens, combinations thereof or any other substances to which the immune system of an egg-producing animal will respond. The critical point in this step is that the immunogen(s) must be capable of inducing an immune state in the egg-producing animal. Although only a single antigen may function as the vaccine for the method of the invention, any mixture of immunogens forming a polyvalent vaccine can also be used, as is well known in the art.

[0045] Alternative vaccines that can be used to immunize the egg-producing animal include genetic vaccines. In particular, any DNA construct (generally consisting of a promoter region and an antigen encoding sequence) will trigger an immune response. Genetic vaccines consist of antigen-coding vectors, fragments of naked DNA, plasmid DNA, DNA-RNA antigens, DNA-protein conjugates, DNA-liposome conjugates, DNA expression libraries, and viral and bacterial DNA delivered to produce an immune response. Methods of DNA delivery include particle bombardment, direct injection, viral vectors, liposomes and jet injection, among others. When applying these delivery methods, much smaller quantities may be necessary and generally result in more persistent antigen production. When using such genetic processes, the preferred method for introducing DNA into avians is through intramuscular injection of the DNA into the breast muscle.

[0046] The vaccine can be either a killed or live-attenuated vaccine and can be administered by any method that elicits an immune response. It is preferred that immunization be accomplished by administering the immunogens through intramuscular injection. The preferred muscle for injection in an avian is the breast muscle. Dosage is preferably 0.05-5 milligrams of the antigenic vaccine. Other methods of administration that can be used include intravenous injection, intraperitoneal injection, intradermal, rectal suppository, aerosol or oral administration. When DNA techniques are used for the immunization process, much smaller quantities are required, generally 1-100 micrograms.

[0047] It can be determined whether the vaccine has elicited an immune response in the egg-producing animal through a number of methods known to those having skill in the art of immunology. Examples of these include enzyme-linked immunosorbent assays (ELISA), tests for the presence of antibodies to the stimulating antigens, and tests designed to evaluate the ability of immune cells from the host to respond to the immunogen. The minimum dosage of immunogen necessary to induce an immune response depends on the vaccination procedure used, including the type of adjuvants and formulation of immunogen(s) used as well as the type of egg-producing animal used as the host.

[0048] In a preferred embodiment, the immunogen used to immunize the target animal is cholecystokinin (CCK) peptide. Isolated CCK peptide has a molecular weight less than 1,500 Daltons. In order to achieve optimal immunogenicity, it is preferred that the CCK peptide be coupled chemically or through recombinant molecular techniques to larger “carrier” molecules. Examples of “carrier” molecules which make a peptide more immunogenic include ovalbumin, bovine gamma globulin (BGG), keyhole limpet hemacyanin (KLH), mouse serum albumin and rabbit serum albumin, among others. Due to its small size, it is preferred that the CCK peptide be conjugated with a carrier protein having a molecular weight of approximately 8,000 Daltons or more in order to form a conjugate of a size capable of eliciting an immune response.

[0049] A preferred method of coupling the CCK peptide to a larger protein carrier to form an immunogen is as follows. The CCK peptide is covalently coupled to a purified carrier protein, such as bovin immunoglobulin G (IgG). Electron-microscopy grade gluteraldehyde [O═CH-(CH₂)₃-CH═O] is preferably used as a homofunctional coupling reagent, where the aldehyde groups form an irreversible bridge between the N-terminal amino group of the peptide and the available amine groups of the protein carrier molecule. This procedure can be applied as a single step wherein the peptide is simultaneously reacted with gluteraldehyde and bovine IgG in the presence of 10 mM sodium acetate, pH 7. Glycine is then added in order to quench any unreacted aldehyde groups that may still be present. The peptide is then dialyzed and a protein assay is performed to determine the concentration of the peptide. The preparation is then preferably aliquoted and stored frozen.

[0050] Once a suitable form of the immunogen is available for immunization, it can then be used to formulate a vaccine. For example, in the case of CCK peptide, the conjugated peptide can be formulated as an adjuvant-based vaccine. This vaccine can then be used to elicit antibody production in a target animal. A typical adjuvant which can be used is Freund's complete adjuvant. If mammals comprise the target animal, then subsequent inoculations should consist of incomplete adjuvant. Other suitable adjuvants include those as referenced in A compendium of vaccine adjuvants and excipients, Vogel, F. R. and Powell, M. F. (1995); In Vaccine Design, The Subunit and Adjuvant Approach, Powell, M. F. and Newman M. J. eds Plenum Press N.Y., as well as others as are known by those having ordinary skill in the art. Amounts and concentration of adjuvant are readily determined by those having ordinary skill in the art.

[0051] In an alternative embodiment, the egg-producing animal is hyperimmunized in order to produce a supranormal level of antibodies. The hyperimmunization procedure is well known in the art, and an example is disclosed in U.S. Pat. No. 4,748,018. Having knowledge of the requirement for developing and maintaining a hyperimmune state, it is within the skill of the art to vary the amount of immunogen administered, depending on the egg-producing animal genera and strain employed, in order to maintain the animal in the hyperimmune state.

[0052] Egg Collection and Processing

[0053] Once the egg-producing animals have been sufficiently immunized to produce an immune response, it is preferred that the eggs from these animals are collected and processed into an administerable egg product.

[0054] Collection and processing of the eggs are the key elements of the present invention. As described in the background, antibodies are produced in the system of an animal during an immune response. These antibodies undergo a maturation process through time in order to develop a mature antibody which binds effectively with an immunogen to neutralize that immunogen. As such, eggs that are collected at different time points after the induction of an immune response will contain antibodies against the same antigen, but the antibodies will be at different stages of maturation.

[0055] The essence of the invention is to mix all eggs which are collected over at least the first 30 week period following immunization. It is preferred that eggs are collected for at least 30 weeks following immunization, and as much as 100 weeks following immunization. It is more preferable that eggs are collected over the 30-60 weeks following immunization.

[0056] In an alternative embodiment, eggs can be collected up until molting. Molting is a process whereby the avians stop laying eggs, and rest until the next laying cycle begins. The decision on when to molt is made by each individual egg producer.

[0057] In a preferred embodiment, eggs collected over the first 30 week period are blended into a single blended lot of egg product. Once eggs laid during at least the first 30 weeks after immunization have been combined into such a blended lot, many options are available for processing. For instance, the eggs themselves can be spray-dried or Immunoglobulin (Ig) can be separated out and purified and then an administerable antibody-containing substance can be produced. Methods for spray-drying and immunoglobulin purification are the subject of several publications and are well known in the art. Prior to administration of the antibody-containing substance, precalculated effective antibody titers, which have previously been determined, are set within these lots so that administerable fractions of the antibody-containing substances contain an effective amount of antibody therein. Once these titers are set, the administerable fraction of antibody-containing substance is administered to a subject animal to passively immunize that subject animal with a more efficacious antibody.

[0058] Antibodies can be isolated and purified from animals by the methods known in the art. For instance, a number of methods for the extraction of antibodies from egg yolk have been described. Polson et al 1985 and Jensenius et al. 1981 successfully used polyethylene glycol and sodium dextran sulfate respectively as protein precipitants in the isolation of pure immunoglobulin from egg yolks. Yokoyama et al. 1992 obtained the water soluble protein fraction after the lipid components were precipitated with an aqueous dispersion of acrylic resins. Lee (U.S. Pat. No. 5,367,054, 1994) describes a high purity and high yield method for isolating and purifying immunoglobulins or fragments thereof from egg yolk by extracting the yolk with a composition containing one or more medium-chain fatty acids.

[0059] Administration

[0060] It is preferred that administration occur by directly feeding blended egg product or any derivative of the blended egg product. The blended egg product can be administered in a variety of ways. For example, the blended egg product can be integrated into a food product. One preferred method for preparing the blended egg product to be incorporated into a food product involves drying the blended egg product into an egg powder. Although various methods are known for drying eggs, spray drying is a preferred method. The process of spray drying eggs is well known in the art. Egg is a natural food ingredients and is non-toxic and safe to those who are not otherwise allergic to eggs.

[0061] The dried egg powder can then be incorporated into drinks in the form of, for example, protein powders, power building drinks, protein supplements and any other nutritional, athlete-associated products. In addition, the egg powder can be used in bake mixes, power bars, candies, cookies, etc. Other examples of egg processing include making an omelet, soft or hard-boiling the egg, baking the egg, or, if desired, the egg can be eaten raw or processed as liquid egg.

[0062] Finally, it is generally known by those having ordinary skill in the art that further separation could provide more potent fractions or elimination of undesirable components, and would allow for other modes of administration such as administering an antibody-containing blend parenterally, subcutaneously, intravenously, intramuscularly, intraperitoneally, intranasally, orally or topically. Such further separation will provide for the ability to make encapsulated products and pharmaceutical compositions with said egg or fraction thereof. It is contemplated that purification could be taken to the level of purified IgY and such purified IgY could then be administered by methods well known in the art.

[0063] It is of significant importance to point out that the egg product of this invention has been shown to be safe, non-toxic, ideal for long term use and has no side effects other than on those animals which are allergic to eggs. The egg product can be orally administered either alone or in combination with drug therapy.

[0064] Alternative Embodiments

[0065] It is further contemplated that one could produce and collect an antibody-containing blend using the process of the invention from other sources such as milk, colostrum, whole blood, plasma and serum of the target animal. It is within the ordinary skill in the art to determine optimum minimum time periods within which to collect milk, serum, etc. to obtain the most efficacious antibody-containing blend.

[0066] The advantageous properties of this invention can be observed by reference to the following examples which illustrate the invention.

EXAMPLES Example 1 Eliciting CCK-8 Antibodies in Eggs

[0067] Methods

[0068] CCK-peptide vaccines were prepared by conjugation of synthetic cholecystokinin (CCK-8) (Fragment 26-33 amide with sulfated tyrosine) to bovine gamma globulin (BGG) using glutaraldehyde. The vaccines were emulsified with Freund's complete adjuvant (1:1) and injected (100 ug CCK) into laying hens. A second injection of the CCK-8 conjugate in Freund's incomplete adjuvant was injected 7 days after primary injection. A second group of control hens did not receive the CCK vaccination. Approximately 2,880 eggs were collected 5 months after the initial injection and the whole eggs were separated into egg yolk and egg white. The egg yolk was spray dried in 8 lots and the antibody titers of the blended spray dried yolk powder were measured.

[0069] Results

[0070] ELISA determinations of the CCK antibody in spray dried egg yolk showed higher end point titers when compared with negative control egg yolk (TABLE 1). Yolks from hens vaccinated with CCK-8 peptide showed an average of 1064 ug/gram in contrast to the negative control egg yolk which contained 3.4 ug/gram specific antibody against CCK-8 peptide. TABLE 1 Analysis of Specific Anti-CCK-8 Antibody Average anti-CCK-8 Antibody (ug/gram Average End Sample yolk) Point Titer Yolk from Hens 1064 394240 Vaccinated with CCK-8 Peptide Conjugate Negative Control 3.4 3379 Yolk

Example 2 Correlation of Blended Lots of CCK AB to Biological Activity in Broiler Chickens

[0071] A total of 6552 one-day-old straight run Ross×Hubbard HyY broiler chicks was used in the experiment. There were 13 treatments with 7 replicates per treatment and 72 birds per replicate. The individual pen was the experimental unit. Blended material from thirty weeks of production was used in this project. A total of six lots were used with a lot being defined as the egg material obtained from approximately 5 weeks of production. Lot 1 was the material obtained from the first 5 weeks of production, Lot 2 was from the second 5 weeks of production, and so on with Lot 6 being from the final 5 weeks (i.e. weeks 25-30) of production. TABLE 2 EXPERIMENTAL TREATMENTS Animals Grams Per Ug Of Antibody Treatment Replicates Per Pen Ton Per Ton Negative 7 72 none 0 Control Lot 1 7 72  46 g/ton 48,195 1050 ug/g 7 72  54 g/ton 56,700 7 72  62 g/ton 65,205 Blended 7 72  71 g/ton 48,195 Lots 7 72 83.5 g/ton  56,700 1-3 680 ug/g 7 72  96 g/ton 65,205 Blended 7 72 220 g/ton 48,195 Lots 7 72 244 g/ton 56,700 4-6 230 ug/g 7 72 281 g/ton 65,205 Blended 7 72 120 g/ton 48,195 Lots 7 72 141 g/ton 56,700 1-6 400 ug/g 7 72 162 g/ton 65,205

[0072] TABLE 3 Weight Gain Treatment 48,195 ug/ton 56,700 ug/ton 65,205 ug/ton Negative 4.566 e Control Lot 1 4.599 cde 4.621 bcde 4.647 abcd Blended Lot 4.596 de 4.630 abcde 4.683 ab 1-3 Blended Lot 4.597 cde 4.640 abcd 4.682 ab 4-6 Blended Lot 4.581 de 4.666 abc 4.694 a 1-6

[0073] TABLE 4 Feed Efficiency Treatment 48,195 ug/ton 56,700 ug/ton 65,205 ug/ton Negative 1.831 f Control Lot 1 1.824 f 1.814 ef 1.792 bcde Blended Lot 1.818 ef 1.802 cdef 1.774 ab 1-3 Blended Lot 1.814 def 1.798 abcd 1.774 abc 4-6 Blended Lot 1.813 def 1.780 abc 1.758 a 1-6

[0074] Results and Conclusions:

[0075] Weight gain and feed efficiency were increased by the administration of an egg product containing a blend of CCK-antibodies in feed. Weight gain and feed efficiency were improved numerically for all doses of CCK antibody blend fed when compared to the negative control. Weight gain and feed efficiency improved numerically within each lot as the dose was increased. The feeding of egg from blending of egg lots to broilers improves weight gain and feed efficiency over the negative control and the positive control (Lot 1). The results of this study indicate statistically that the blending of CCK antibody should include at least 30 weeks of production material before being used commercially in order to achieve the most benefit to performance. 

We claim:
 1. A method for increasing efficacy of an antibody in a subject comprising: A. inducing an immune response in an egg-producing animal with at least one immunogen; B. collecting eggs from the egg-producing animal for a predetermined period of time following induction of said immune response; C. processing all of the collected eggs together to form a blended mix; and D. administering to the subject a portion of the blended mix which contains an effective titer of said antibody.
 2. The method of claim 1 wherein the predetermined period of time comprises at least 30 weeks.
 3. The method of claim 2 wherein the predetermined period of time comprises 30 to 60 weeks.
 4. The method of claim 1 wherein the immunogen is selected from the group consisting of proteins, enzymes, bacteria, viruses, protozoa, fungi, cellular antigens, allergens and combinations thereof.
 5. The method of claim 4 wherein the immunogen comprises cholecystokinin.
 6. The method of claim 1 wherein the egg-producing animal comprises an avian.
 7. The method of claim 6 wherein the avian is selected from the group consisting of fowls, turkeys, ducks and geese.
 8. An antibody-containing substance comprising an effective titer of at least one antibody, said antibody comprising all antibody structures produced within an animal over a predetermined period of time following induction of an immune response in said animal.
 9. The antibody-containing substance of claim 8 wherein the animal comprises an egg-producing animal.
 10. The antibody-containing substance of claim 9 further comprising an egg product.
 11. The antibody-containing substance of claim 10 wherein the egg product comprises an effective amount of egg obtained from a blended mix of all eggs produced by one or more egg-producing animal for at least 30 weeks following induction of the immune response.
 12. An antibody-containing substance comprising an effective titer of at least one antibody, said antibody-containing substance comprising an egg product obtained from a blended mix of all eggs produced over a predetermined period of time by one or more egg-producing animals which have been immunized with at least one immunogen.
 13. The antibody-containing substance of claim 12 wherein the predetermined period of time comprises at least 30 weeks.
 14. The antibody-containing substance of claim 13 wherein the predetermined period of time comprises 30 to 60 weeks.
 15. The antibody-containing substance of claim 12 wherein the immunogen comprises an antigen selected from the group consisting of proteins, enzymes, bacteria, viruses, protozoa, fungi, cellular antigens, allergens and combinations thereof.
 16. The antibody-containing substance of claim 15 wherein the immunogen comprises cholecystokinin.
 17. The antibody-containing substance of claim 12 wherein the egg-producing animal comprises an avian.
 18. The antibody-containing substance of claim 17 wherein the avian is selected from the group consisting of fowls, turkeys, ducks and geese. 